EP0409473A2 - A process for obtaining the gangliosides from nervous tissue of mammalians and for the treatment of psychoorganic diseases with said material compositons - Google Patents
A process for obtaining the gangliosides from nervous tissue of mammalians and for the treatment of psychoorganic diseases with said material compositons Download PDFInfo
- Publication number
- EP0409473A2 EP0409473A2 EP90307575A EP90307575A EP0409473A2 EP 0409473 A2 EP0409473 A2 EP 0409473A2 EP 90307575 A EP90307575 A EP 90307575A EP 90307575 A EP90307575 A EP 90307575A EP 0409473 A2 EP0409473 A2 EP 0409473A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- gangliosides
- alcohol
- exchange material
- ion exchange
- process according
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- 150000002270 gangliosides Chemical class 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims abstract description 30
- 239000000463 material Substances 0.000 title claims abstract description 26
- 230000008569 process Effects 0.000 title claims abstract description 24
- 238000011282 treatment Methods 0.000 title claims description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims description 11
- 201000010099 disease Diseases 0.000 title claims description 9
- 239000000203 mixture Substances 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000005342 ion exchange Methods 0.000 claims abstract description 18
- 239000011877 solvent mixture Substances 0.000 claims abstract description 18
- 239000003495 polar organic solvent Substances 0.000 claims abstract description 14
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 11
- 239000003513 alkali Substances 0.000 claims abstract description 7
- 238000005192 partition Methods 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims abstract description 6
- 238000010521 absorption reaction Methods 0.000 claims abstract description 5
- 239000000706 filtrate Substances 0.000 claims abstract description 5
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 4
- 238000001914 filtration Methods 0.000 claims abstract description 4
- 150000002576 ketones Chemical class 0.000 claims abstract description 4
- 239000003960 organic solvent Substances 0.000 claims abstract description 4
- 238000004440 column chromatography Methods 0.000 claims abstract description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 208000020401 Depressive disease Diseases 0.000 claims description 10
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 claims description 10
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 9
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 8
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Natural products CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 8
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 claims description 8
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 claims description 7
- 208000024714 major depressive disease Diseases 0.000 claims description 7
- 230000001537 neural effect Effects 0.000 claims description 7
- 230000007996 neuronal plasticity Effects 0.000 claims description 7
- 230000003982 neuronal uptake Effects 0.000 claims description 7
- 229960002748 norepinephrine Drugs 0.000 claims description 7
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 claims description 7
- 239000002899 monoamine oxidase inhibitor Substances 0.000 claims description 6
- XNWFRZJHXBZDAG-UHFFFAOYSA-N 2-METHOXYETHANOL Chemical compound COCCO XNWFRZJHXBZDAG-UHFFFAOYSA-N 0.000 claims description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical group CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 5
- 201000003104 endogenous depression Diseases 0.000 claims description 5
- 230000000144 pharmacologic effect Effects 0.000 claims description 5
- 238000011084 recovery Methods 0.000 claims description 5
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims description 4
- 241000283690 Bos taurus Species 0.000 claims description 4
- 239000002671 adjuvant Substances 0.000 claims description 4
- 229910021529 ammonia Inorganic materials 0.000 claims description 4
- 208000035755 Psychosomatic disease Diseases 0.000 claims description 3
- 239000001913 cellulose Substances 0.000 claims description 3
- 229920002678 cellulose Polymers 0.000 claims description 3
- 238000001704 evaporation Methods 0.000 claims description 3
- 230000008020 evaporation Effects 0.000 claims description 3
- 210000000278 spinal cord Anatomy 0.000 claims description 3
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 claims description 2
- 208000019022 Mood disease Diseases 0.000 claims description 2
- 208000012826 adjustment disease Diseases 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 201000003995 melancholia Diseases 0.000 claims description 2
- 208000015238 neurotic disease Diseases 0.000 claims description 2
- 238000007738 vacuum evaporation Methods 0.000 claims description 2
- 210000001638 cerebellum Anatomy 0.000 claims 2
- 239000012736 aqueous medium Substances 0.000 claims 1
- 230000000063 preceeding effect Effects 0.000 claims 1
- 125000003944 tolyl group Chemical group 0.000 claims 1
- 239000000935 antidepressant agent Substances 0.000 description 15
- 210000001519 tissue Anatomy 0.000 description 15
- 229940005513 antidepressants Drugs 0.000 description 12
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 10
- 229920005654 Sephadex Polymers 0.000 description 10
- 239000012507 Sephadex™ Substances 0.000 description 10
- 239000002904 solvent Substances 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 6
- 230000001430 anti-depressive effect Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- 150000001298 alcohols Chemical class 0.000 description 5
- 210000002569 neuron Anatomy 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 230000001225 therapeutic effect Effects 0.000 description 5
- 230000006399 behavior Effects 0.000 description 4
- 210000003169 central nervous system Anatomy 0.000 description 4
- ZPUCINDJVBIVPJ-LJISPDSOSA-N cocaine Chemical compound O([C@H]1C[C@@H]2CC[C@@H](N2C)[C@H]1C(=O)OC)C(=O)C1=CC=CC=C1 ZPUCINDJVBIVPJ-LJISPDSOSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 125000005629 sialic acid group Chemical group 0.000 description 4
- 150000003408 sphingolipids Chemical class 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 229940123685 Monoamine oxidase inhibitor Drugs 0.000 description 3
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000001154 acute effect Effects 0.000 description 3
- 230000003044 adaptive effect Effects 0.000 description 3
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 150000002402 hexoses Chemical group 0.000 description 3
- 229960004592 isopropanol Drugs 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 230000036651 mood Effects 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 230000009182 swimming Effects 0.000 description 3
- 230000004797 therapeutic response Effects 0.000 description 3
- WWUZIQQURGPMPG-UHFFFAOYSA-N (-)-D-erythro-Sphingosine Natural products CCCCCCCCCCCCCC=CC(O)C(N)CO WWUZIQQURGPMPG-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 231100000111 LD50 Toxicity 0.000 description 2
- SUHOOTKUPISOBE-UHFFFAOYSA-N O-phosphoethanolamine Chemical compound NCCOP(O)(O)=O SUHOOTKUPISOBE-UHFFFAOYSA-N 0.000 description 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 230000003542 behavioural effect Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 229940106189 ceramide Drugs 0.000 description 2
- 229960003920 cocaine Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 229930182478 glucoside Natural products 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- YPFDHNVEDLHUCE-UHFFFAOYSA-N propane-1,3-diol Chemical compound OCCCO YPFDHNVEDLHUCE-UHFFFAOYSA-N 0.000 description 2
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- WWUZIQQURGPMPG-KRWOKUGFSA-N sphingosine Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)CO WWUZIQQURGPMPG-KRWOKUGFSA-N 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000008733 trauma Effects 0.000 description 2
- UOCLXMDMGBRAIB-UHFFFAOYSA-N 1,1,1-trichloroethane Chemical compound CC(Cl)(Cl)Cl UOCLXMDMGBRAIB-UHFFFAOYSA-N 0.000 description 1
- NUKLDTSTOVGVDB-UHFFFAOYSA-N 1,1-dichloroethane;1,2-dichloroethane Chemical compound CC(Cl)Cl.ClCCCl NUKLDTSTOVGVDB-UHFFFAOYSA-N 0.000 description 1
- KFUSEUYYWQURPO-UHFFFAOYSA-N 1,2-dichloroethene Chemical group ClC=CCl KFUSEUYYWQURPO-UHFFFAOYSA-N 0.000 description 1
- LRYZPFWEZHSTHD-HEFFAWAOSA-O 2-[[(e,2s,3r)-2-formamido-3-hydroxyoctadec-4-enoxy]-hydroxyphosphoryl]oxyethyl-trimethylazanium Chemical class CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](NC=O)COP(O)(=O)OCC[N+](C)(C)C LRYZPFWEZHSTHD-HEFFAWAOSA-O 0.000 description 1
- ZNQVEEAIQZEUHB-UHFFFAOYSA-N 2-ethoxyethanol Chemical compound CCOCCO ZNQVEEAIQZEUHB-UHFFFAOYSA-N 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 208000020925 Bipolar disease Diseases 0.000 description 1
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 description 1
- 206010010144 Completed suicide Diseases 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- HCYAFALTSJYZDH-UHFFFAOYSA-N Desimpramine Chemical compound C1CC2=CC=CC=C2N(CCCNC)C2=CC=CC=C21 HCYAFALTSJYZDH-UHFFFAOYSA-N 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 241000722985 Fidia Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- GVGLGOZIDCSQPN-PVHGPHFFSA-N Heroin Chemical compound O([C@H]1[C@H](C=C[C@H]23)OC(C)=O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4OC(C)=O GVGLGOZIDCSQPN-PVHGPHFFSA-N 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- ZAGRKAFMISFKIO-UHFFFAOYSA-N Isolysergic acid Natural products C1=CC(C2=CC(CN(C2C2)C)C(O)=O)=C3C2=CNC3=C1 ZAGRKAFMISFKIO-UHFFFAOYSA-N 0.000 description 1
- 206010024870 Loss of libido Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- SQVRNKJHWKZAKO-PFQGKNLYSA-N N-acetyl-beta-neuraminic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)O[C@H]1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-PFQGKNLYSA-N 0.000 description 1
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 description 1
- SUHQNCLNRUAGOO-UHFFFAOYSA-N N-glycoloyl-neuraminic acid Natural products OCC(O)C(O)C(O)C(NC(=O)CO)C(O)CC(=O)C(O)=O SUHQNCLNRUAGOO-UHFFFAOYSA-N 0.000 description 1
- FDJKUWYYUZCUJX-UHFFFAOYSA-N N-glycolyl-beta-neuraminic acid Natural products OCC(O)C(O)C1OC(O)(C(O)=O)CC(O)C1NC(=O)CO FDJKUWYYUZCUJX-UHFFFAOYSA-N 0.000 description 1
- FDJKUWYYUZCUJX-KVNVFURPSA-N N-glycolylneuraminic acid Chemical compound OC[C@H](O)[C@H](O)[C@@H]1O[C@](O)(C(O)=O)C[C@H](O)[C@H]1NC(=O)CO FDJKUWYYUZCUJX-KVNVFURPSA-N 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- DPWPWRLQFGFJFI-UHFFFAOYSA-N Pargyline Chemical compound C#CCN(C)CC1=CC=CC=C1 DPWPWRLQFGFJFI-UHFFFAOYSA-N 0.000 description 1
- GMZVRMREEHBGGF-UHFFFAOYSA-N Piracetam Chemical compound NC(=O)CN1CCCC1=O GMZVRMREEHBGGF-UHFFFAOYSA-N 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 206010039897 Sedation Diseases 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- 208000032140 Sleepiness Diseases 0.000 description 1
- 206010041349 Somnolence Diseases 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 208000007271 Substance Withdrawal Syndrome Diseases 0.000 description 1
- 206010042458 Suicidal ideation Diseases 0.000 description 1
- 241001661355 Synapsis Species 0.000 description 1
- 231100000644 Toxic injury Toxicity 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 230000008649 adaptation response Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 150000001338 aliphatic hydrocarbons Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000000164 antipsychotic agent Substances 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 206010003549 asthenia Diseases 0.000 description 1
- -1 benzene or toluene Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 208000028683 bipolar I disease Diseases 0.000 description 1
- 208000025307 bipolar depression Diseases 0.000 description 1
- DWBRWHDWQZWHIC-UHFFFAOYSA-N butan-2-ol;2-methylpropan-1-ol Chemical compound CCC(C)O.CC(C)CO DWBRWHDWQZWHIC-UHFFFAOYSA-N 0.000 description 1
- FFGPTBGBLSHEPO-UHFFFAOYSA-N carbamazepine Chemical compound C1=CC2=CC=CC=C2N(C(=O)N)C2=CC=CC=C21 FFGPTBGBLSHEPO-UHFFFAOYSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical group 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 description 1
- 150000001783 ceramides Chemical class 0.000 description 1
- 210000003710 cerebral cortex Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000005607 chanvre indien Nutrition 0.000 description 1
- 244000261228 chanvre indien Species 0.000 description 1
- ZPEIMTDSQAKGNT-UHFFFAOYSA-N chlorpromazine Chemical compound C1=C(Cl)C=C2N(CCCN(C)C)C3=CC=CC=C3SC2=C1 ZPEIMTDSQAKGNT-UHFFFAOYSA-N 0.000 description 1
- 231100000160 chronic toxicity Toxicity 0.000 description 1
- 230000007665 chronic toxicity Effects 0.000 description 1
- 230000002060 circadian Effects 0.000 description 1
- 239000000549 coloured material Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 229960002069 diamorphine Drugs 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 206010013663 drug dependence Diseases 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 239000012156 elution solvent Substances 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 229940093476 ethylene glycol Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000002339 glycosphingolipids Chemical class 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- BCGWQEUPMDMJNV-UHFFFAOYSA-N imipramine Chemical compound C1CC2=CC=CC=C2N(CCCN(C)C)C2=CC=CC=C21 BCGWQEUPMDMJNV-UHFFFAOYSA-N 0.000 description 1
- 229960004801 imipramine Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000012442 inert solvent Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 206010022437 insomnia Diseases 0.000 description 1
- ULYZAYCEDJDHCC-UHFFFAOYSA-N isopropyl chloride Chemical compound CC(C)Cl ULYZAYCEDJDHCC-UHFFFAOYSA-N 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 208000020442 loss of weight Diseases 0.000 description 1
- ZAGRKAFMISFKIO-QMTHXVAHSA-N lysergic acid Chemical compound C1=CC(C2=C[C@H](CN([C@@H]2C2)C)C(O)=O)=C3C2=CNC3=C1 ZAGRKAFMISFKIO-QMTHXVAHSA-N 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001730 monoaminergic effect Effects 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 1
- SNMVRZFUUCLYTO-UHFFFAOYSA-N n-propyl chloride Chemical compound CCCCl SNMVRZFUUCLYTO-UHFFFAOYSA-N 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 150000002812 neutral glycosphingolipids Chemical class 0.000 description 1
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 description 1
- 239000002664 nootropic agent Substances 0.000 description 1
- 231100001143 noxa Toxicity 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 229960001779 pargyline Drugs 0.000 description 1
- 208000027232 peripheral nervous system disease Diseases 0.000 description 1
- 208000033808 peripheral neuropathy Diseases 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 238000011458 pharmacological treatment Methods 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 229950004354 phosphorylcholine Drugs 0.000 description 1
- PYJNAPOPMIJKJZ-UHFFFAOYSA-N phosphorylcholine chloride Chemical compound [Cl-].C[N+](C)(C)CCOP(O)(O)=O PYJNAPOPMIJKJZ-UHFFFAOYSA-N 0.000 description 1
- 229960004526 piracetam Drugs 0.000 description 1
- 230000001242 postsynaptic effect Effects 0.000 description 1
- 230000009290 primary effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 235000013772 propylene glycol Nutrition 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000036280 sedation Effects 0.000 description 1
- 230000000862 serotonergic effect Effects 0.000 description 1
- 229940076279 serotonin Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 231100000456 subacute toxicity Toxicity 0.000 description 1
- 208000011117 substance-related disease Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 210000003568 synaptosome Anatomy 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/02—Acyclic radicals, not substituted by cyclic structures
- C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
- C07H15/10—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical containing unsaturated carbon-to-carbon bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/26—Psychostimulants, e.g. nicotine, cocaine
Definitions
- This invention is concerned with a process for obtaining mixtures of gangliosides from mammalian nervous tissue and for their use in the treatment of psycho-organic diseases.
- compositions which include fractions or mixtures of gangliosides extracted from mammalian nervous tissue, having pharmacological activity and being useful as adjuvants for the treatment of diseases of the mood which require the use of monoamine oxidase inhibitors or neuronal uptake blockers of noradrenaline or serotonin. Likewise, they are useful for treating psycho-organic diseases which require the recovery of damaged neurons and stimulation of neuronal plasticity.
- gangliosides comprise one of the three groups constituting the sphingolipids family, complex lipids which are present in great amounts in the nervous tissues (brain, cerebral cortex, spinal cord, etc.), and only in small amounts in the adipose tissue.
- the sphingolipids contain three building blocks: a fatty acid molecule (C18-C20) linked by means of peptide bond to an aminoalcohol, generally named "sphingosine” (of which no less than 30 different species are known), making up units called “ceramides", for example:- is a ceramide of stearic acid and sphingosine.
- the third building block arises from substitution of the hydroxy group at the C1 position.
- lipids of this type are the "neutral glycosphingolipids", wherein the alcoholic function at the C1 position is blocked by a glucoside bond to a neutral carbohydrate residue (galactose, glucose, etc.). These compounds are present in high quantities in neural tissue and to a lesser extent in non-neural tissue.
- a third group of compounds included under the donomination of sphingolipids are the "acid glycosphingolipids" usually called “gangliosides", wherein the 1-alcohol function is also blocked by a glucoside bond with an oligosaccharide molecule carrying sialic acid.
- gangliosides the "acid glycosphingolipids" usually called “gangliosides”
- N-acetyl neuraminic acid is usually found in human gangliosides, N-glycolyl neuraminic acid also being quite common.
- gangliosides More than 20 different types have been identified, which differ in the quantity and in the relative positions of the hexose and the sialic acid residues.
- Gangliosides containing only one fragment of sialic acid are called monosialogangliosides; if they contain 2, they are called disialogangliosides; if they contain 3, they are called trisialogangliosides, and if they contain 4, they are called tetrasialogangliosides. They are indicated as GM, GD, GT and GQ, respectively.
- a numerical suffix and a letter are added. The numerical suffix is equal to 5-n, wherein n is the number of neutral hexose units.
- the letters (generally a, b) indicate isomeric positions of the sialic acids fragments. These fragments have different Rf values in thin layer chromatography.
- gangliosides are effective for inducing functional changes in synaptosomes and plastic changes in neurons cultured in vitro , existing numerous works which demonstrate this action [ B. Maggio, F.A. Cumar, R. Caputto, Biochem-Biophys Acta 1981, 650, 69-87, and M.C. Byrne, R.W. Ledeen, F.J. Raisen, G. Yorkee and J.R. Scalafani, J. Neurochem. 1983 41 1214-1222 ].
- gangliosides facilitate functional neuronal plasticity.
- Basic experimental evidence shows that gangliosides facilitate a series of behavioural responses connected with neuronal plasticity [ H. Rahmann, in Learning and memory: mechanisms of information storage in the nervous system, ed. H. Matthies Pergamon Press, N. Y. 1986, p.235-245 ].
- the purification known until now consists basically in precipitation or dialysis. Precipitation always leaves some active product in solution and also drags out impurities, thus increasing the number of purification steps needed. Dialysis is long and expensive and also leads to losses in the yield of active product.
- An object of the present invention is to provide an improved process for obtaining gangliosides from mammalian nervous tissue for the treatment of psycho-organic diseases, which require neuronal plasticity recovery or the use of monoamine-oxidase inhibitors or neuronal uptake blockers of noradrenaline or serotonine.
- the present invention resides in a process for obtaining gangliosides from mammalian nervous tissue, comprising the steps of:-
- the gangliosides are recovered in step (d) from the ion exchange material by eluting them with said solvent mixture, distilling the solvent mixture, and extracting the residue with a solvent mixture including water, (C1-C4) alcohol(s) and non-polar organic solvent(s), the solvent mixture extract being subjected to the partition or absorption column chromotography of purifying step (e).
- a solvent mixture including water, (C1-C4) alcohol(s) and non-polar organic solvent(s)
- the alkali is preferably a volatile alkali such as ammonia or an amine.
- the recovering step (h) is preferably effected by eluting the column with a mixture of water, (C1-C4) alcohol(s) and non-polar organic solvent(s) using solvent mixtures of increasing polarity, and recovering the fractions which contain the gangliosides.
- the fractions which contain gangliosides are subjected to evaporation to evaporate the solvent mixture, and the residue that contains the extracted gangliosides is recovered.
- the ion exchange material of step (b) is preferably a dextran- or cellulose-based material (eg DEAE Sephadex or DEAE cellulose, dimethylaminoethycellulose) preferably applied at a ratio of 0.8-1.5 g for each 0.1 g of gangliosides to be adsorbed.
- a dextran- or cellulose-based material eg DEAE Sephadex or DEAE cellulose, dimethylaminoethycellulose
- Another object of the present invention is to obtain an active substance composition having pharmacological activity useful as an adjuvant in the treatment of disorders of the mood ascribed to the block of neuronal uptake noradrenaline or serotonine or in diseases treated with monoamine-oxidase inhibitors, and even useful for treating psychosomatic diseases which require the re-establishment of neuronal plasticity.
- the present invention provides a composition having pharmacological activity useful as an adjuvant in the treatment of mood disorders such as unipolar endogenous depression, bipolar endogenous depression, melancholia or reactive depression treated with neuronal uptake blockers of noradrenaline or serotonine or in diseases treated with monoamino-oxidase inhibitors, or the treatment of psychosomatic diseases which require the recovery of neuronal, said composition comprising, in an administrable form, gangliosides extracted from mammalian nervous tissue in a pharmaceutically acceptable vehicle.
- the method for obtaining gangliosides according to the present invention avoids the previously mentioned disadvantageous prior art steps, and proposes an extraction and partition between a non-polar organic solvent phase and an aqueous alcoholic phase [ J. Folch, M. Lees and G.B. Sloane-Stanley, J. Biol. Chem., 226 , 497 (195)] followed by a purification in an ion exchange modified cellulose (DEAE Sephadex), and a selective elution thereof, under such conditions that the yield is higher than the previously described methods and the costs are lower.
- the ratio of gangliosides to ion exchange material can be significantly higher than those described by Takashi Momol, Susumo Ando and Yoshitaka Nagai in Biochim. Biophys., Acta 441 448/97 (1976).
- the solvents for elution proposed do not contain salts, thus avoiding desalination steps (dialysis). Therefore, our production method results in a process which is simpler than those used until now, simplifying some stages and eliminating the more complicated ones. These stages can be performed in a bathwise manner, avoiding the use of columns which demand more time.
- the exchange material used preferably DEAE Sephadex
- DEAE Sephadex is left in such a condition that it can be regenerated by washing it with dilute acetic acid solution, afterwards with water, and finally with methanol-water mixture and then reused.
- the eluted product is finally purified by chromatography on silica gel or any other suitable solid support, eg a natural or synthetic silicate such as HYFLOW or CELITE. Due to its nature, our method allows a yield which is higher than the methods described by Lars Svennerholm in J. Neurochem. 10 , 613 (1963) or in Belgium Patent (Fidia) 843695 and Patent 7540170, thereby ensuring that the final product will have a higher quality.
- aromatic hydrocarbons such as benzene or toluene
- aliphatic hydrocarbons such as n-heptane or n-hexane, or mixtures of isomers of hexanes
- chlorinated solvents such as dichloromethane, chloroform, 1,1-dichloroethane 1,2-dichloroethane, dichlorethylene, trichloroethane, 2-chloropropane, 1-chloropropane, etc. can be used as non-polar organic solvents.
- the alcohols used to form the aqueous alcoholic phase can be monohydric alcohols (eg C1 -C4 alcohols such as methanol, ethanol, n-propanol, 2-propanol, n-butanol, isobutanol (2-butanol), methyl cellosolve or ethyl-cellosolve ) or dihydric alcohols (eg C2 -C3 alcohols such as ethyleneglycol, 1,3-dihydroxypropane, or 1,2-dihydroxypropane ).
- monohydric alcohols eg C1 -C4 alcohols such as methanol, ethanol, n-propanol, 2-propanol, n-butanol, isobutanol (2-butanol), methyl cellosolve or ethyl-cellosolve
- dihydric alcohols eg C2 -C3 alcohols such as ethyleneglycol, 1,3-
- Example 1 is repeated, but using 265 ml of dichloromethane as the non-polar organic solvent, and 200 ml of methylcellosolve (ethyleneglycol monomethylether) as the alcohol.
- Example 1 is repeated, but using 265 ml of 1,2-dichlorethane as the non-polar organic solvent, and 130 ml of n-butanol as the alcohol.
- Example 1 is repeated, but using 320 ml of chloroform as the non-polar organic solvent, and 260 ml of methanol as the alcohol.
- Example 1 is repeated, but using dichloroethylene as the inert solvent, and isobutanol as the alcohol.
- the aqueous alcoholic phase obtained according to Example 1, 2, 3, 4 or 5 is treated with DEAE Sephadex at the ratio of 0.8 g for each 120 mg of gangliosides to be absorbed in a 1-litre beaker, for 20 hours under mild stirring. Then, it is vacuum filtered using a Buchner-type funnel, discarding the filtrate. Afterwards, the DEAE Sephadex is washed over the filter with a solvent containing a carbonyl group (acetone).
- Gangliosides are eluted from the DEAE Sephadex by washing the latter with a mixture (56: 35: 3: 2 by vol.) of dichloroethane: isopropanol: water: methylamine (40%), in a batchwise manner for 20 minutes, the water: methylamine (40%) corresponding to a 40%w/w solution of methylamine in water.
- the liquors are gathered and filtered.
- Example 6 the DEAE Sephadex is washed with a mixture (58.3: 33.8: 4.17: 3.6 by vol.) of chloroform: methanol: water: ammonia, the water-ammonia system corresponding to a 25%w/w solution of ammonia in water.
- the elution solvent obtained according to Example 6, 7 or 8 is removed from the extracted gangliosides by vacuum evaporation at 40°C in a rotary evaporation until dryness is achieved. The residue is redissolved in a mixture (74.5: 23: 2.5 by vol.) of chloroform: methanol: distilled water.
- a silica gel column is prepared using the solvent described in the previous step.
- the solvent is drained off up to dryness.
- the residue is taken up in a small amount of the same solvent and the obtained ganglioside solution is seeded in the column. It is then eluted with an additional amount of the same solvent, continuing with increasing polar mixtures of chloroform: methanol: distilled water.
- the term “neuronal plasticity” indicates the ability of the nervous system to change or modify its accquired responses to enviromental stimuli, exogenous or endogenous stimulations, or traumatic and/or toxic injuries.
- the plasticity of the central nervous system is the condition thereof to permit the repair of damage tissue, give organic base for the memory process and respond to therapeutics with behavioural changes.
- Disorders of mood or behaviour comprise the major endogenous depression and the bipolar depression or maniac-depressive diseases which are mainly characterized by changes in character, comprising feelings of deep sadness, mental slowness and loss of concentration, pessimistic worries and an intense sense of minus value. These physical changes are accompanied by various organic signs, such as a characteristic insomnia, anorexia, asthenia, loss of weight and libido diminution and interruption of the hormonal circadian cycles. All these signs and symptoms are accompanied by suicidal tendencies, suicide being a certain and relatively high risk probability (15%).
- the forced swimming test is an example of this methodology.
- the animal under experimentation is placed in a container filled with water under controlled temperature, where it can swim but lacks any possibility to escape. After several minutes of forced swimming, the animal accepts the impossibility of escaping and ceases its efforts to escape, letting itself float. When this test is repeated 10 days later, the animal does not try to swim, thus accepting the lack of escape, with its immobility. If in the interval between both tests, the animal has received treatment with an antidepressant, the animal swims vigorously, trying to find an outlet.
- gangliosides together with antidepressant drugs to animals under experimentation, reduces the dose of antidepressant and time of treatment required to obtain the intended effect.
- This potentiation would be related to the facilitation of the plastic phenomena of the central nervous system produced by gangliosides [ V.A. Molina, E.A. Keller and O.A. Orsingher, European J. Pharmacol., 1989, 160 , 247-252).
- mice and rats were studied: Acute, subacute and chronic toxicity in mice and rats and presence of pyrogen or histamine-like substances. These last two assays were "negative", indicating the purity of the mixture. As regards toxicity, the lethal dose 50% (LD50) obtained was about 2.5 g/kg, indicating a wide therapeutic index.
- LD50 lethal dose 50%
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Psychiatry (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Pain & Pain Management (AREA)
- Ophthalmology & Optometry (AREA)
- Immunology (AREA)
- Virology (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
- (a) extracting the nervous tissue using at least one (C₁-C₄) alcohol and at least one non-polar organic solvent,
- (b) filtering the mixture and adding water to the filtrate to produce an aqueous alcoholic phase,
- (c) adsorbing gangliosides present in said aqueous alcoholic phase on an ion exchange material, and then washing the ion exchange material with a ketone,
- (d) recovering the gangliosides from the ion exchange material using a solvent mixture including water, (C₁-C₄) alcohol(s), organic solvent(s) and alkali(es),
- (e) purifying the material recovered from the ion exchange material by partition or absorption column chromatography, and
- (f) recovering the extracted gangliosides from the chromatography column.
Description
- This invention is concerned with a process for obtaining mixtures of gangliosides from mammalian nervous tissue and for their use in the treatment of psycho-organic diseases.
- Furthermore, this invention provides compositions which include fractions or mixtures of gangliosides extracted from mammalian nervous tissue, having pharmacological activity and being useful as adjuvants for the treatment of diseases of the mood which require the use of monoamine oxidase inhibitors or neuronal uptake blockers of noradrenaline or serotonin. Likewise, they are useful for treating psycho-organic diseases which require the recovery of damaged neurons and stimulation of neuronal plasticity.
- It is known that gangliosides comprise one of the three groups constituting the sphingolipids family, complex lipids which are present in great amounts in the nervous tissues (brain, cerebral cortex, spinal cord, etc.), and only in small amounts in the adipose tissue.
- The sphingolipids contain three building blocks: a fatty acid molecule (C₁₈-C₂₀) linked by means of peptide bond to an aminoalcohol, generally named "sphingosine" (of which no less than 30 different species are known), making up units called "ceramides", for example:-
- The third building block arises from substitution of the hydroxy group at the C₁ position.
-
- Another group of complex lipids of this type, are the "neutral glycosphingolipids", wherein the alcoholic function at the C₁ position is blocked by a glucoside bond to a neutral carbohydrate residue (galactose, glucose, etc.). These compounds are present in high quantities in neural tissue and to a lesser extent in non-neural tissue.
- A third group of compounds included under the donomination of sphingolipids, are the "acid glycosphingolipids" usually called "gangliosides", wherein the 1-alcohol function is also blocked by a glucoside bond with an oligosaccharide molecule carrying sialic acid. Of these acids, N-acetyl neuraminic acid is usually found in human gangliosides, N-glycolyl neuraminic acid also being quite common.
- More than 20 different types of gangliosides have been identified, which differ in the quantity and in the relative positions of the hexose and the sialic acid residues.
- Gangliosides containing only one fragment of sialic acid are called monosialogangliosides; if they contain 2, they are called disialogangliosides; if they contain 3, they are called trisialogangliosides, and if they contain 4, they are called tetrasialogangliosides. They are indicated as GM, GD, GT and GQ, respectively. A numerical suffix and a letter are added. The numerical suffix is equal to 5-n, wherein n is the number of neutral hexose units. The letters (generally a, b) indicate isomeric positions of the sialic acids fragments. These fragments have different Rf values in thin layer chromatography.
-
- GM:₁ monosialoganglioside with four units of neutral hexose (n=4), see the TABLE hereinafter.
- GD1a: disialoganglioside, wherein n=4, and has specific distributions of sialic acid fragments.
- GD1b: disialoganglioside, wherein n=4 and exhibits isomerism in connection to the position of the sialic acid fragment, with respect to GD1a.
- The importance of gangliosides in the transmission of the nerve impulse through the synapsis and the immunity reactions of tissue and cell-to-cell recognition is known.
- Moreover, taking into account the high proportion of gangliosides in relation to other sphingolipids in the membranes of the neurons, it has been established that they accomplish an important function in the structure and function thereof. Exogenously added, gangliosides are effective for inducing functional changes in synaptosomes and plastic changes in neurons cultured in vitro, existing numerous works which demonstrate this action [ B. Maggio, F.A. Cumar, R. Caputto, Biochem-Biophys Acta 1981, 650, 69-87, and M.C. Byrne, R.W. Ledeen, F.J. Raisen, G. Yorkee and J.R. Scalafani, J. Neurochem. 1983 41 1214-1222 ]. These effects, which may be observed in a tissue culture, are also verifiable when they are injected in vivo into the animal under experimentation. Thus, several groups of researchers in different parts of the world have demonstrated that exogenous administration of gangliosides facilitates the repair of neuronal tissue which has been injured by different types of noxa [ H.A. Tilson, G.J. Harry, K. Nanry, P.M. Hudson, J.S. Hong, J of Neurosc. Res. 1988, 19, 88-93 ]. However, all works above mentioned limit the field of action of gangliosides to anatomical repair of the injured cell and, therefore, restrict the potential medical use thereof to structural alterations of neurons, cause by trauma, metabolic or toxic effect such as different peripheral neuropathies, caused by diabetes, alcoholism, anti-neoplastic drugs, trauma, etc, and therefore can be said to restrict the therapeutic field of action of gangliosides [ R. Abraham, D.M. Levy and R.M. Abraham, Diabetes Research 1988, 7, 129-135 ].
- It has now been proved that it is possible to apply gangliosides to facilitate functional neuronal plasticity. Basic experimental evidence shows that gangliosides facilitate a series of behavioural responses connected with neuronal plasticity [ H. Rahmann, in Learning and memory: mechanisms of information storage in the nervous system, ed. H. Matthies Pergamon Press, N. Y. 1986, p.235-245 ].
- The proposal that the mixture of gangliosides is an experimental medicine which has been developed for the treatment of afflictions related to behaviour is an hypothesis confirmed by definite experimental evidence.
- As regards the extraction methods of gangliosides known up to the moment, they use numerous steps for obtaining a crude product, including the employment of both peroxide-forming solvents as well as different types of ethers. The peroxides can react with the double bonds reducing the yield of the final products.
- The purification known until now consists basically in precipitation or dialysis. Precipitation always leaves some active product in solution and also drags out impurities, thus increasing the number of purification steps needed. Dialysis is long and expensive and also leads to losses in the yield of active product.
- An object of the present invention is to provide an improved process for obtaining gangliosides from mammalian nervous tissue for the treatment of psycho-organic diseases, which require neuronal plasticity recovery or the use of monoamine-oxidase inhibitors or neuronal uptake blockers of noradrenaline or serotonine.
- Thus, in one aspect, the present invention resides in a process for obtaining gangliosides from mammalian nervous tissue, comprising the steps of:-
- (a) extracting the nervous tissue using at least one (C₁-C₄) alcohol and at least one non-polar organic solvent,
- (b) filtering the mixture and adding water to the filtrate to produce an aqueous alcoholic phase,
- (c) adsorbing gangliosides present in said aqueous alcoholic phase on an ion exchange material (preferably an anionic exchange material), and then washing the ion exchange material with a ketone,
- (d) recovering the gangliosides from the ion exchange material using a solvent mixture including water, (C₁-C₄) alcohol(s), organic solvent(s) and alkali(es),
- (e) purifying the material recovered from the ion exchange material by partition or absorption column chromatography, and
- (f) recovering the extracted gangliosides from the chromatography column.
- Preferably, the gangliosides are recovered in step (d) from the ion exchange material by eluting them with said solvent mixture, distilling the solvent mixture, and extracting the residue with a solvent mixture including water, (C₁-C₄) alcohol(s) and non-polar organic solvent(s), the solvent mixture extract being subjected to the partition or absorption column chromotography of purifying step (e).
- The alkali is preferably a volatile alkali such as ammonia or an amine.
- The recovering step (h) is preferably effected by eluting the column with a mixture of water, (C₁-C₄) alcohol(s) and non-polar organic solvent(s) using solvent mixtures of increasing polarity, and recovering the fractions which contain the gangliosides.
- Preferably, the fractions which contain gangliosides are subjected to evaporation to evaporate the solvent mixture, and the residue that contains the extracted gangliosides is recovered.
- The ion exchange material of step (b) is preferably a dextran- or cellulose-based material (eg DEAE Sephadex or DEAE cellulose, dimethylaminoethycellulose) preferably applied at a ratio of 0.8-1.5 g for each 0.1 g of gangliosides to be adsorbed.
- Another object of the present invention is to obtain an active substance composition having pharmacological activity useful as an adjuvant in the treatment of disorders of the mood ascribed to the block of neuronal uptake noradrenaline or serotonine or in diseases treated with monoamine-oxidase inhibitors, and even useful for treating psychosomatic diseases which require the re-establishment of neuronal plasticity.
- Thus, in another aspect of the present invention, the present invention provides a composition having pharmacological activity useful as an adjuvant in the treatment of mood disorders such as unipolar endogenous depression, bipolar endogenous depression, melancholia or reactive depression treated with neuronal uptake blockers of noradrenaline or serotonine or in diseases treated with monoamino-oxidase inhibitors, or the treatment of psychosomatic diseases which require the recovery of neuronal, said composition comprising, in an administrable form, gangliosides extracted from mammalian nervous tissue in a pharmaceutically acceptable vehicle.
- The method for obtaining gangliosides according to the present invention, avoids the previously mentioned disadvantageous prior art steps, and proposes an extraction and partition between a non-polar organic solvent phase and an aqueous alcoholic phase [ J. Folch, M. Lees and G.B. Sloane-Stanley, J. Biol. Chem., 226, 497 (195)] followed by a purification in an ion exchange modified cellulose (DEAE Sephadex), and a selective elution thereof, under such conditions that the yield is higher than the previously described methods and the costs are lower.
- The ratio of gangliosides to ion exchange material (eg DEAE Sephadex) can be significantly higher than those described by Takashi Momol, Susumo Ando and Yoshitaka Nagai in Biochim. Biophys., Acta 441 448/97 (1976). The solvents for elution proposed do not contain salts, thus avoiding desalination steps (dialysis). Therefore, our production method results in a process which is simpler than those used until now, simplifying some stages and eliminating the more complicated ones. These stages can be performed in a bathwise manner, avoiding the use of columns which demand more time. Moreover, and concerning the economy of the process, the exchange material used (preferably DEAE Sephadex) is left in such a condition that it can be regenerated by washing it with dilute acetic acid solution, afterwards with water, and finally with methanol-water mixture and then reused. By washing the DEAE Sephadex that retains the gangliosides, with solvents having a carbonyl group, a substantial proportion of coloured material dragged out as an impurity is eliminated.
- The eluted product is finally purified by chromatography on silica gel or any other suitable solid support, eg a natural or synthetic silicate such as HYFLOW or CELITE. Due to its nature, our method allows a yield which is higher than the methods described by Lars Svennerholm in J. Neurochem. 10, 613 (1963) or in Belgium Patent (Fidia) 843695 and Patent 7540170, thereby ensuring that the final product will have a higher quality.
- In the first stage, that is to say, extraction, aromatic hydrocarbons, such as benzene or toluene, or aliphatic hydrocarbons, such as n-heptane or n-hexane, or mixtures of isomers of hexanes, or chlorinated solvents such as dichloromethane, chloroform, 1,1-dichloroethane 1,2-dichloroethane, dichlorethylene, trichloroethane, 2-chloropropane, 1-chloropropane, etc. can be used as non-polar organic solvents. The alcohols used to form the aqueous alcoholic phase can be monohydric alcohols ( eg C₁ -C₄ alcohols such as methanol, ethanol, n-propanol, 2-propanol, n-butanol, isobutanol (2-butanol), methyl cellosolve or ethyl-cellosolve ) or dihydric alcohols ( eg C₂ -C₃ alcohols such as ethyleneglycol, 1,3-dihydroxypropane, or 1,2-dihydroxypropane ).
- The following examples are given solely for the purpose of illustrating the different ways that exist to carry out the present invention.
- 100 g of homogenized bovine encephalic matter are placed in a 2 liter beaker, and 196 ml of isopropanol are added with stirring to produce a homogenous paste to which 265 ml of toluene are added with continued stirring. The beaker is covered and stirring is continued for 2 hours. Then, it is filtered under vacuum.
- To the clear liquid derived from the previous filtration, water is added at the ratio of about 22 ml for each 100 ml of filtrate. After smooth stirring to secure mixing, it is left standing for 24 hours. The phases are separated and the organic solvent phase is discarded, leaving the aqueous alcoholic phase available for subsequent operations.
-
- Example 1 is repeated, but using 265 ml of dichloromethane as the non-polar organic solvent, and 200 ml of methylcellosolve (ethyleneglycol monomethylether) as the alcohol.
- Example 1 is repeated, but using 265 ml of 1,2-dichlorethane as the non-polar organic solvent, and 130 ml of n-butanol as the alcohol.
- Example 1 is repeated, but using 320 ml of chloroform as the non-polar organic solvent, and 260 ml of methanol as the alcohol.
- Example 1 is repeated, but using dichloroethylene as the inert solvent, and isobutanol as the alcohol.
- The aqueous alcoholic phase obtained according to Example 1, 2, 3, 4 or 5 is treated with DEAE Sephadex at the ratio of 0.8 g for each 120 mg of gangliosides to be absorbed in a 1-litre beaker, for 20 hours under mild stirring. Then, it is vacuum filtered using a Buchner-type funnel, discarding the filtrate. Afterwards, the DEAE Sephadex is washed over the filter with a solvent containing a carbonyl group (acetone).
- Gangliosides are eluted from the DEAE Sephadex by washing the latter with a mixture (56: 35: 3: 2 by vol.) of dichloroethane: isopropanol: water: methylamine (40%), in a batchwise manner for 20 minutes, the water: methylamine (40%) corresponding to a 40%w/w solution of methylamine in water. The liquors are gathered and filtered.
- In a modification Example 6, the DEAE Sephadex is washed with a mixture (60: 36: 0.85: 2.2 by vol.) of methylene chloride: n-butanol: water: methylamine 40%.
- In a further modification of Example 6, the DEAE Sephadex is washed with a mixture (58.3: 33.8: 4.17: 3.6 by vol.) of chloroform: methanol: water: ammonia, the water-ammonia system corresponding to a 25%w/w solution of ammonia in water.
- The elution solvent obtained according to Example 6, 7 or 8 is removed from the extracted gangliosides by vacuum evaporation at 40°C in a rotary evaporation until dryness is achieved. The residue is redissolved in a mixture (74.5: 23: 2.5 by vol.) of chloroform: methanol: distilled water.
- Afterwards, a silica gel column is prepared using the solvent described in the previous step.
- The solvent is drained off up to dryness. The residue is taken up in a small amount of the same solvent and the obtained ganglioside solution is seeded in the column. It is then eluted with an additional amount of the same solvent, continuing with increasing polar mixtures of chloroform: methanol: distilled water.
- 10 ml fractions are collected, and spectrophotometrically screened for phospholipids and gangliosides according to Lars Svennarholm in Biochim. Biophys. Acta 24, 604 (1957) using a color reaction. Constant flow is maintained in the column. The fractions richer in gangliosides are selected and then gathered. The gathered fractions are filtered and evaporated to dryness under vacuum in a rotary evaporator. They are taken up again in distilled water, and the pH is adjusted to 7.8 using diluted solutions of hydrochloric acid and sodium hydroxide and then are lyophilized. These operations yield between 90 and 180 mg of gangliosides per 100 g fresh tissue.
Usual experimental yield: 150-170 mg. - This yield is at least three times higher than the yield achieved in Belgian Patent No. 843695.
- Today the term "neuronal plasticity" indicates the ability of the nervous system to change or modify its accquired responses to enviromental stimuli, exogenous or endogenous stimulations, or traumatic and/or toxic injuries. In short, the plasticity of the central nervous system is the condition thereof to permit the repair of damage tissue, give organic base for the memory process and respond to therapeutics with behavioural changes.
- The clearest example of plastic or adaptive reactions in response to a pharmacological treatment and connected with a therapeutic effect is undoubtedly the one which takes place during the treatment with antidepressant drugs.
- Disorders of mood or behaviour comprise the major endogenous depression and the bipolar depression or maniac-depressive diseases which are mainly characterized by changes in character, comprising feelings of deep sadness, mental slowness and loss of concentration, pessimistic worries and an intense sense of minus value. These physical changes are accompanied by various organic signs, such as a characteristic insomnia, anorexia, asthenia, loss of weight and libido diminution and interruption of the hormonal circadian cycles. All these signs and symptoms are accompanied by suicidal tendencies, suicide being a certain and relatively high risk probability (15%).
- Many of the drugs used to treat endogeneous depression exert deep effects on the metabolism of a group of chemical substances, that is, the monoamines: noradrenaline and serotonine, which act as neurotransmitters in the Central Nervous System. The more conspicuous effect is to block the neuronal uptake mechanisms for amines. When the neuronal uptake is blocked or impaired, the effect of these neurotransmitters on postsynaptic or effector neurons is enhanced, being both potentiated in magnitude and prolonged in time.
- Over the period 1960-1975, the pharmacodynamic basis for the therapeutic action of antidepressants was ascribed to this blocking of uptake. However, various inconsistencies demonstrated that the phenomenon was more complex. Thus, for example, although the blocking of the monoamine inactivation is immediate upon the administration of the drug, the clinical therapeutic response is only achieved after a period of about 15 days of continuous treatment. Furthermore, several drugs that also cause the inhibition of the uptake, such as amphetamines or cocaine are not antidepressant and, even though they produce behavioural activation when administered in an acute manner, they lack clinical efficacy as antidepressants. On the contrary, the antidepressants produce sedation and somnolence at the beginning of the treatment.
- Later studies allow a more complex theory on the mechanism of action of the antidepressants to be conceived. To a primary effect related to the increase in the monoaminergic neurotransmission, there follows an adaptive or plastic response of certain neuronal circuits which involve a decrease in the number of receptors betaadrenergic and serotonergic. These plastic responses, which take more than two weeks to be completed, would be the organic substrate to the therapeutic response to antidepressants.
- On the basis of these new theories, the methods for experimental searches of antidepressants have varied. Formerly, the acute affect connected in principle with the evaluation of the effectiveness as blockers of the inactivation mechanism was investigated. At present, instead, there is an attempt to evaluate the effect of an extended treatment of the animal over certain behaviours, thus seeking the results of plastic changes which took place following the treatment.
- The forced swimming test is an example of this methodology. The animal under experimentation is placed in a container filled with water under controlled temperature, where it can swim but lacks any possibility to escape. After several minutes of forced swimming, the animal accepts the impossibility of escaping and ceases its efforts to escape, letting itself float. When this test is repeated 10 days later, the animal does not try to swim, thus accepting the lack of escape, with its immobility. If in the interval between both tests, the animal has received treatment with an antidepressant, the animal swims vigorously, trying to find an outlet. This test is very sensitive and allows the gradual obtention of responses (time of swimming), which are proportional to the administered doses (dose-response curve) or to the duration of treatment (time-response curve) [ R.D. Porsolt, G. Anton, N. Blavet and M. Jalfre, European J. Pharmacol., 1978, 47, 379-383 ].
- The administration of gangliosides together with antidepressant drugs to animals under experimentation, reduces the dose of antidepressant and time of treatment required to obtain the intended effect. This potentiation would be related to the facilitation of the plastic phenomena of the central nervous system produced by gangliosides [ V.A. Molina, E.A. Keller and O.A. Orsingher, European J. Pharmacol., 1989, 160, 247-252).
- These and other similar experiments lead to the conclusion that the administration of a ganglioside mixture can improve the depressed patient's therapeutic response to the known antidepressant. It is proposed then:
- 1. The use of a ganglioside mixture (extracted, for example, from bovine, porcine, equine, ovine encephalon), containing mainly GM1, GD1a, GD1b, and GT1b to facilitate the therapeutic effect of antidepressant drugs, such as dimethylimipramine, imipramine, nortiptiline, maproptiline, pargyline, etc.
- 2. The above mentioned use may be applicable to reduce the dosage of antidepressant or reduce the time necessary between the beginning of the treatment and the therapeutic effect of the antidepressant.
- 3. The concomitant use of gangliosides with other pharmacological therapeutics, causing adaptive effects on the central nervous system, such as:
anti-epilectic drugs: diphenylhydantione, carbamezepine,
nootropic drugs:piracetam
antipsychotic drugs: chloropromazine, etc. - 4. The use of the ganglioside mixture in the treatment of drug dependence syndromes to facilitate the adaptive response, during the period of drug withdrawal such as: morphine, heroin, marihuana, cocaine, amphetamines and diethylamine derivates of lysergic acid, etc.
- Before using said ganglioside mixture, the following variables were studied: Acute, subacute and chronic toxicity in mice and rats and presence of pyrogen or histamine-like substances. These last two assays were "negative", indicating the purity of the mixture. As regards toxicity, the lethal dose 50% (LD₅₀) obtained was about 2.5 g/kg, indicating a wide therapeutic index.
Claims (18)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AR314371 | 1989-07-12 | ||
AR89314371A AR241751A1 (en) | 1989-07-12 | 1989-07-12 | Proceeding for obtaining mixtures from gangliosides for treatment of character and psycho-organic disorders. |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0409473A2 true EP0409473A2 (en) | 1991-01-23 |
EP0409473A3 EP0409473A3 (en) | 1992-04-29 |
Family
ID=3478704
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19900307575 Ceased EP0409473A3 (en) | 1989-07-12 | 1990-07-11 | A process for obtaining the gangliosides from nervous tissue of mammalians and for the treatment of psychoorganic diseases with said material compositons |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP0409473A3 (en) |
JP (1) | JPH03135991A (en) |
AR (1) | AR241751A1 (en) |
BR (1) | BR9003470A (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993008808A2 (en) * | 1991-11-08 | 1993-05-13 | Fidia S.P.A | Use of gangliosides in the preparation of pharmaceuticals which can improve the progonosis of disorders due to psychotropics and drugs abuse |
WO2010062197A1 (en) * | 2008-11-25 | 2010-06-03 | Eduard Nekrasov | Dairy product and process |
CN109705176A (en) * | 2019-01-23 | 2019-05-03 | 苏州纳微科技股份有限公司 | The isolation and purification method of one boar gangliosides |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2320760A1 (en) * | 1975-08-13 | 1977-03-11 | Fidia Spa | Mixtures of mono-, di-, tri- and tetra-sialo-gangliosides - for treatment of disorders due to conduction of nervous stimuli of the central and peripheral nervous systems |
EP0150712A2 (en) * | 1984-01-04 | 1985-08-07 | Bioiberica, S.A. | Method for the obtention of a glycosphingolipid complex |
EP0319890A1 (en) * | 1987-12-09 | 1989-06-14 | Crinos Industria Farmacobiologica S.p.A. | A process for the preparation of monosialoganglioside |
-
1989
- 1989-07-12 AR AR89314371A patent/AR241751A1/en active
-
1990
- 1990-07-11 EP EP19900307575 patent/EP0409473A3/en not_active Ceased
- 1990-07-12 JP JP2185154A patent/JPH03135991A/en active Pending
- 1990-07-12 BR BR909003470A patent/BR9003470A/en unknown
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2320760A1 (en) * | 1975-08-13 | 1977-03-11 | Fidia Spa | Mixtures of mono-, di-, tri- and tetra-sialo-gangliosides - for treatment of disorders due to conduction of nervous stimuli of the central and peripheral nervous systems |
EP0150712A2 (en) * | 1984-01-04 | 1985-08-07 | Bioiberica, S.A. | Method for the obtention of a glycosphingolipid complex |
EP0319890A1 (en) * | 1987-12-09 | 1989-06-14 | Crinos Industria Farmacobiologica S.p.A. | A process for the preparation of monosialoganglioside |
Non-Patent Citations (1)
Title |
---|
CHEMICAL ABSTRACTS, vol. 85, no. 21, 22nd November 1976, page 214, abstract no. 155928n, Columbus, Ohio, US; T. MOMOI et al.: "High resolution preparative column chromatographic system for gangliosides using DEAE-Sephadex and a new porous silica, Iatrobeads" & BIOCHIM. BIOPHYS. ACTA 1976, 441(3), 488-97 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993008808A2 (en) * | 1991-11-08 | 1993-05-13 | Fidia S.P.A | Use of gangliosides in the preparation of pharmaceuticals which can improve the progonosis of disorders due to psychotropics and drugs abuse |
WO1993008808A3 (en) * | 1991-11-08 | 1993-06-10 | Fidia Spa | Use of gangliosides in the preparation of pharmaceuticals which can improve the progonosis of disorders due to psychotropics and drugs abuse |
WO2010062197A1 (en) * | 2008-11-25 | 2010-06-03 | Eduard Nekrasov | Dairy product and process |
CN109705176A (en) * | 2019-01-23 | 2019-05-03 | 苏州纳微科技股份有限公司 | The isolation and purification method of one boar gangliosides |
Also Published As
Publication number | Publication date |
---|---|
BR9003470A (en) | 1991-08-27 |
AR241751A1 (en) | 1992-12-30 |
JPH03135991A (en) | 1991-06-10 |
EP0409473A3 (en) | 1992-04-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE3750785T2 (en) | Oligosaccharides of heparin with an affinity for factors of cell growth. | |
KR0160104B1 (en) | Method for treatment of senile dementia | |
DE69830251T2 (en) | LONG PENTRAXIN PTX3 CONTAINING PHARMACEUTICAL COMPOSITIONS | |
EP0049847A2 (en) | Alpha-amylase inactivator, process for its preparation, composition containing said inactivator, and this inactivator for the regulation of the blood sugar level | |
US4707469A (en) | Gangliosides mixture, useful as a therapeutical tool for eliminating painful effects or peripheral neuropathies | |
DE60126416T2 (en) | TREATMENT OF ARTHRITIS AND CORRESPONDING COMPOSITIONS | |
DE2549680C2 (en) | ||
DE60124532T2 (en) | NEW POLYPEPTIDES FROM BEEKE POISON AND METHOD FOR THEIR USE | |
CN1799552A (en) | Single sialic acid tetrahexose ganglioside preparation method | |
CN1302811A (en) | Compound (I), extraction process and its application in preparing medicine to cure acute ischemic cerebrovascular disease | |
EP0409473A2 (en) | A process for obtaining the gangliosides from nervous tissue of mammalians and for the treatment of psychoorganic diseases with said material compositons | |
Tilson et al. | Ganglioside interactions with the dopaminergic system of rats | |
US5190925A (en) | Use of gangliosides in the treatment of autonomic dysfunction in Chagas' disease | |
DE3624816A1 (en) | METHOD FOR PRODUCING MONOSIAL GANGLIOSIDE | |
DE60109279T2 (en) | PHARMACEUTICAL COMPOSITIONS WITH OLIGOSACCHARIDES AND THEIR PREPARATION | |
DE3787971T2 (en) | ACTIVE PRINCIPLE, ISOLATED FROM HAIGEWEWE. | |
DE2605576C3 (en) | Process for isolating the proteases papain, chimopapain, lysozyme and proteinase X from the milk sap of Carica papya and use of the isolated proteases for the production of sterilized and lyophilized orthopedic, neurosurgical or ophthalmological preparations | |
JP3126162B2 (en) | Manufacturing method of nerve cell activator | |
DE1959933A1 (en) | Process for the preparation of drugs containing ganglioside sulphates with antibiotic effects by extracting material of animal origin | |
DE3881484T2 (en) | Optically active products of 20,21-dinoreburnamenin derivatives, processes for their preparation, their use as medicaments and pharmaceutical compositions containing them. | |
DE68911049T2 (en) | Therapeutic use of the isopropyl ester derivative of monosialogangliosides in diseases of the nervous system, accompanied by inflammation. | |
Date | The isolation of some oligosaccharides from the urine of pregnant and lactating women | |
EP2533787B1 (en) | Use of isorhamnetin triglycosides | |
IT9022520A1 (en) | PROCESS FOR OBTAINING GANGLIOSIDES FROM THE NERVOUS TISSUE OF MAMMALS AND FOR THE TREATMENT OF PSYCHOORGANIC DISORDERS WITH COMPOUNDS OF SUCH MATERIALS. | |
JPH0386822A (en) | Nervous cell alternation-repairing agent |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE |
|
RBV | Designated contracting states (corrected) |
Designated state(s): AT BE CH DE DK ES FR GB GR IT LI NL SE |
|
PUAL | Search report despatched |
Free format text: ORIGINAL CODE: 0009013 |
|
AK | Designated contracting states |
Kind code of ref document: A3 Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE |
|
17P | Request for examination filed |
Effective date: 19921026 |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: LABORATORIOS BETA S.A. |
|
17Q | First examination report despatched |
Effective date: 19941011 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED |
|
18R | Application refused |
Effective date: 19951017 |